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Catalog # HK318

Elafin (Trappin-2), Human, ELISA kit

778.001,282.00

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Elafin is an epithelial proteinase inhibitor also known under various other names such as Skin-derived Anti leukoproteinase (SKALP) and Elastase-Specific Inhibitor (ESI). Elafin belongs to the Trappin gene family and was given the systematic name Trappin-2 in a recent classification. The Trappin family is defined by a N-terminal transglutaminase substrate domain and a C-terminal four disulphide core. Trappins have been suggested to play a role in the regulation of inflammation and in protection against tissue damage in stratified epithelia. Elafin is an inhibitor of leukocyte elastase and proteinase-3 and in addition it is a substrate for transglutaminases. The protein is constitutively expressed in various epithelia including those of hair follicles, oesophagus, vagina and oral cavity. Elafin is not present in normal human skin but is strongly induced during inflammation as in psoriasis and wound healing. The full-length protein is translated as a 12.3 kDa protein of 117 amino acids termed pre-elafin or Trappin-2. Cleavage of the signal peptide yields a mature protein with a molecular mass of 9.9 kDa. The 9.9 kDa secreted protein is the major form found in culture medium. In skin extracts a 6 kDa form comprising the 57 most C-terminal amino acids is present, which is the form of elafin originally described by Wiedow et al. In serum both the 9.9 and the 6 kDa form appear to be present. In urine only the short 6 kDa form is found. The Hycult biotech ELISA is not suitable to measure the short 6 kDa form. Elafin has been used as a quantitative marker for disease activity in severe psoriasis during treatment with cyclosporin. In serum/plasma of healthy individuals approximately 10-50 ng/ml elafin is present. Elafin shows a 10-fold increase in concentration during psoriasis. The determination of Elafin is not disturbed by the presence of the short 6 kDa form.

Application
The human Elafin ELISA kit is to be used for the in vitro quantitative determination of human Elafin in serum, plasma and cell culture supernatant samples. The ELISA has no cross reactivity with Elafin/SKALP (6 kDa form).

Principle
The human elafin ELISA is a ready-to-use solid-phase enzyme-linked immunosorbent assay based on the sandwich principle with a working time of 3½ hours. The efficient format of 2 plates with twelve disposable 8-well strips allows free choice of batch size for the assay. Samples and standards are captured by a solid bound specific antibody. Biotinylated tracer antibody will bind to captured human elafin. Streptavidin-peroxidase conjugate will bind to the biotinylated tracer antibody. Streptavidin-peroxidase conjugate will react with the substrate, tetramethylbenzidine (TMB). The enzyme reaction is stopped by the addition of oxalic acid. The absorbance at 450 nm is measured with a spectrophotometer. A standard curve is obtained by plotting the absorbance (linear) versus the corresponding concentrations of the human elafin standards (log). The human elafin concentration of samples, which are run concurrently with the standards, can be determined from the standard curve.

Product type
Assays
Amount
1 x 96 det., 2 x 96 det.
Standard range
878-10,000 pg/ml
Detection level
878 pg/ml
Working volume
100 µl/well
Species
Human
Cross reactivity
Bovine – Yes, Horse – Yes, Rabbit – Yes
Storage and stability
Product should be stored at 4 °C. Under recommended storage conditions, product is stable for at least six months.
Precautions
For research use only. Not for use in or on humans or animals or for diagnostics. It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. Hycult Biotech is not responsible for any patent infringements that might result with the use of or derivation of this product.
Disease
Autoimmunity, Gastroenterology
Safety Data Sheet Assay Manual
HK318
402 kb

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